Team:BGU Israel/Part Composite

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Composite Part
BBa_K2938015 - Cry11Aa (Strep tag) + deGFP Device

Link: Cry11Aa (Strep) +deGFP Device

In our project, we wanted to express the Bacillus thuringiensis subsp. israelensis (Bti) toxin subunits in Serratia Marcescens 274. In order to do so, we planned and constructed three initial, “minimal”, polycistronic plasmids, in order to combine them into one final plasmid containing all the genes present in these plasmids.

We haven’t been able to construct our final plasmid yet, and this is why we carried out experiments with our initial “minimal” plasmids.

Our best composite part is one of our initial plasmid devices, and the one that showed greatest toxicity yet.


Other Basic parts we added:
In our project we used the Bacillus thuringiensis subsp. israelensis (Bti) toxin subunits. Some of these toxin genes were used by previous iGEM teams and projects, but had various point mutations throughout the DNA sequence compared to the published sequences. That is why we added these basic parts again.
Also, we designed and added commonly used tags (HA, Strep and His) to our subunits in order to be able to detect them using Western Blot analysis using anti-tags antibodies.
The parts containing the tags appear in our designed plasmids (devices) that we used and presented during our project.

We also found some changes between our tags and tags that were already in the registry, so we added our versions of the tags. Also, we added the terminator sequence that we used in all of our devices.

The Composite part is a polycistronic plasmid containing the following basic parts:

  1. PR1 promoter - our best basic part, a promoter that can be used in a polycistronic plasmid and shows exceptional translation rates.
  2. Cry11Aa - a Bti toxic subunit, which has larvicidal activity against mosquito larvae.
  3. RBS - between the different genes, in order to translate the many genes on the mRNA.
  4. deGFP - a GFP marker.
  5. T500 terminator - a terminator originating from the pBEST plasmid.

Cry4Ba Plasmid Cry4Ba Plasmid
Cry11Aa + P20 Plasmid Cry11Aa + P20 Plasmid
Final Plasmid Final Plasmid

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